DNA-binding profiling of human hormone nuclear receptors via fluorescence correlation spectroscopy in a cell-free system.
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Kobayashi T, Kodani Y, Nozawa A, Endo Y, Sawasaki T
DNA-binding profiling of human hormone nuclear receptors via fluorescence correlation spectroscopy in a cell-free system.
FEBS Lett. 2008 Aug 6;582(18):2737-44. doi: 10.1016/j.febslet.2008.07.003. Epub 2008 Jul 11.
- PubMed ID
- 18619963 [ View in PubMed]
- Abstract
The nuclear hormone receptors (NHRs), a family of transcription factors, bind directly to the hormone response elements (HREs) to regulate gene expression. In this study, we describe a comprehensive NHR-HRE profiling analysis with a new high-throughput DNA binding assay system utilizing wheat germ cell-free protein production and fluorescence correlation spectroscopy (FCS). This approach revealed NHR binding to natural response elements and new heterodimeric NHR-HRE bindings. We analyzed 408 possible binding combinations between 34 human NHRs and 12 different HREs, and identified 205 NHR-HRE binding combinations, 124 of which have not been previously reported. Thus, this study provides a novel biochemical classification of the human NHRs, as well as describing a novel approach to the large-scale analysis of DNA-protein interactions.